Fig 1: Ribosomal collisions at PTCs decrease sup-tRNA efficacy.a Nuclease-resistant disomes from Calu3 expressing W1282X-CFTR or WT-CFTR. Different ribosomal fractions (monosomes—black, disomes— green, and polysomes—blue) were separated through sucrose gradients (horizontal gray triangle designates the sucrose gradient) and probed with GCN1 and ZNF598 antibodies. The numbers under the blots denote normalized protein band (NP) normalized to the loaded protein measured as total protein amount. b Simultaneous probing with CFTR and ZNF598 antibodies of Calu3 cells expressing WT-CFTR or W1282X-CFTR. Cells expressing W1282X-CFTR and not treated with sup-tRNA (untreated) produced a shorter CFTR variant (marked with *). Treatment with sup-tRNA completely abolished the strong ZNF598 signal. Numbers under the blots denote the normalized protein band (NP) ratioed to the loaded protein measured as total protein amount. These experiments serve as a quality check for disomes (a) or collision markers (b) and hence, were performed as single experiments. Source data are provided as a Source Data file.